Development of Quantitative Real-time PCR and Loop-mediated Isothermal Amplification (LAMP) Assays for Detection of Microsporidium seriolae

被引：0

作者：

Mekata, Tohru ^{[1
]}

Satoh, Jun ^{[1
]}

Nakayasu, Chihaya ^{[1
]}

Ishii, Yuji ^{[2
]}

Harakawa, Shogo ^{[2
]}

Kawakami, Hidemasa ^{[2
]}

Yanagi, Soetsu ^{[3
]}

机构：

[1] Japan Fisheries Res & Educ Agcy, Aquaculture Res Dept, Nansei Branch, Fisheries Technol Inst, Minamiise, Mie 5160193, Japan

[2] Ehime Res Inst Agr Forestry & Fisheries, Ehirne Fisheries Res Ctr, Uwajima, Ehime 7980087, Japan

[3] Kagoshirna Prefecture Fisheries Promot Sect, Kagoshima 8908577, Japan

来源：

FISH PATHOLOGY | 2021年 / 56卷 / 02期

关键词：

Microsporidium seriolae; real-time PCR; LAMP; BEKO DISEASE; VISUAL DETECTION; FISH; YELLOWTAIL; SINGLE;

D O I：

暂无

中图分类号：

S9 [水产、渔业];

学科分类号：

0908 ;

摘要：

Beko disease, caused by the Microsporidium seriolae infection, has been a problem in yellowtail aquaculture in western Japan. In recent years, severe cases of this disease have been confirmed, resulting in a significant decrease in product value due to mass mortality and residual cysts. Only polymerase chain reaction (PCR) assays have been reported so far as a detection method for the disease. In this study, quantitative real-time PCR (qPCR) and loop-mediated isothermal amplification (LAMP) methods have been optimized for M. seriolae detection in order to establish a more sensitive and rapid diagnosis. Target regions for each detection method were selected based on the nucleotide sequences obtained by the gene analysis of cysts in diseased fish. Primer sets for the qPCR and LAMP methods were designed, and the gene amplification efficiency of each method was evaluated. The results showed that the newly developed qPCR method could detect 1.4 copies of the target gene, and the LAMP method detected 100 copies within 15 minutes. In this study, the newly developed qPCR and LAMP assays were shown to be rapid and highly sensitive methods for quantitative detection of M. seriolae.

引用

页码：53 / 61

页数：9

共 18 条

[1] Single and nested polymerase chain reaction assays for the detection of Microsporidium seriolae (Microspora), the causative agent of 'Beko' disease in yellowtail Seriola quinqueradiata [J].